Multiplexed Surface Plasmon Resonance Imaging for Protein Biomarker Analysis
The reliable detection of ligand and analyte binding is of significant importance for the field of medical diagnostics. Recent advances in proteomics and the rapid expansion in the number of identified protein biomarkers enhance the need for reliable techniques for their identification in complex samples. Surface plasmon resonance imaging (SPRi) provides label-free detection of this binding process in real-time. This chapter details the fabrication of an SPR imaging instrument and its use in analyzing molecular binding interactions with the use of a high-density microfluidic SPRi chip, capable of multiplexed analysis as well as various immobilization chemistries. Controlled recovery of bound biomarkers is demonstrated to enable their identification using mass spectrometry. Finally, activated leukocyte cell adhesion molecule (ALCAM), a protein biomarker associated with a variety of cancers, is identified from human crude cell lysates using the microfluidic surface plasmon resonance imaging (SPRi) instrument.
Key wordsSurface plasmon resonance (SPR) imaging Microfluidic arrays Poly(dimethylsiloxane) PDMS HeLa cells Biomarkers Mass spectrometry Self-assembled monolayers
The authors would like to thank Dr. Leonard Foster for his advice and continued support, Dr. Leroy Hood and Christopher Lausted for their contributions to the microfluidic device design, Anders Riss Kristensen for his technical assistance with HeLa cell culture and Dr. Robert Parker for technical assistance with mass spectrometry analysis. Microfabrication was performed in the UBC Nanolab.
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